CA2256303A1 - The use of mmp inhibitors for the treatment of ocular angiogenesis - Google Patents

Abstract

The present invention relates to the use of matrix metalloproteinase inhibitors,preferably those which display specihcity for matrix metalloproteinases-2 or 9, in the treatment or prevention of ocular angiogenesis.

Description

.

BACKGROUND OF THE INVENTION
The present invention relates to the use of matrix meta cpr,tu;.,ase inh;~;tcl,~preferably those which display specificit~ for matrix " ,etallopr~,t~inases-2 or 9 in the treatment or prevention of ocular ang;ogenesis i.e macular degeneration and diabetic retinopathy.
Aberrant angiogenesis occurs in several different disease areas including cancerrheumatoid arthritis and psoriasi~. An~ic3snesis also occurs in various eye d;~a-~s such as diabetic reffnopathy and age rebted macubr degeneldlion (ARMD). Taken together 2.4 million individuals are affected by these two ocular indications (0.7 million for pr~ 'erdti~e diabebc retinopdU,~ and 1.7 million for angiogenic ARMD). Laser ph k -~a3L'aticn is a highhy effective 15 therapy for prc 'eralive diabetic r~tinopdU"~ but has side effects primarily a variable bss of penpheral and night vision. Currently there is no therapy capable of preventing or adequately treating ARMD.
Comp'.: 'icns of diabetes is the major cause of h ~dness in patents under 50 years of age In diabetic retinopdU,~ new blood ~essels overlying the retina develop exhibit i"c~ased 20 vascular permeability micro-anurysms and her"o~l,age resulting in damage to the neural retina.
The current therapy for diabetic ,~tinop~U.~ is to phclx a3u ~ the retina outside the macula so that the blood vessels are destroyed and cannot further pr~ '; rat_. This ~edt",er,l has a success rate of appro~i-,-a~ 95% but has side effects primarily a variable bss of pe-i~heidl and night vision. The incdence of d;~t~ i related ocular an_ 3~nesis is 25 000 casesfyear.
Age-related macular degeneration (ARMD) is the leading cause of legal t ,dl,ess and low vision in the elderly. This disorder ,~:pr~sentj an advanced and pdU,c!c3 - stage of the aging process that occurs in all eyes. Ag~related macular degeneration exists in a ~dry~ or ~wer fomm.
The ~dry~ or atrophic type causes gradual deterioration of vision over 10 to 20 years. The ~wer or exudative type causes a rapid and v~iJesp.~ad loss of central vision due to leakage of blood or 30 fluid from abnormal newly fom ed blood vessels (i.e. choroidal neovascular membranes) prol,f~rdLing beneath the macula. Sinoe this process occurs in the macula there is a major impact on acuity and oentral vision. Unlike diabetes related t ,dness patients with ARMD are frequent~y healthy in other respects and the loss of vision can convert an active independenl individual into one totally depende"l on others. In ARMD one eye usually shows symptoms first 35 and patients are faced with several years of anguish waiting for the likeiy loss of sight in the other eye. There is no effecbve way to treat or prevent this con 'iticn. Pl,o' ~ag l~ation therapy cannot be used sinoe it destroys the ph t )~e~l,lui~ and would cause t ~lness if applied to the macula. A category of ARMD cor~er~d to have a high risk of dcvl ~ F lg the exudative type is distinguishable by clinical examination. S~ce the prevalence of this category of ARMD doubles 40 with every decade after 50 and the aging of the popu~a'ions of the dcl~eloped countries the suffering and health care costs of this cor, 'i~ n are likely to grow Jldlll_L--~Iy. In a US
popu'a~n study in males over the age d thirty the prevalence of exudative disease was 0.5%

CA 022~6303 1998-12-17 but the prevalence of the high risk condltion, (prime candldates for prophylactic therapy) was 9% (Bessler et al, Arch Opthalmol, 107, 847(1989). In the over 70 age group prevalence of the high risk condition rose to 26%. This is a ma~or unmet medical need.
The present inventor has now dlscovered that matrlx metalloprotelnase lnhlbltors, preferably those whlch dlsplay speclflcity for matrix metalloproteinases-2 or 9, are useful for the treatment or preventlon of ocular anglogenesis, l.e.
macular degeneration and diabetlc retinopathy.
SUMMARY OF THE INVENTION
The present invention relates to a pharmaceutlcal composltion, preferably, an intraorbltol compositlon for treating or preventlng ocular anglogenesis, lncludlng macular degeneratlon and dlabetlc retlnopathy, by lnhlbltlng a matrix metalloprotelnase ln a mammal, preferably a human, comprlsing a matrlx metalloprotelnase inhibltlng effective amount of a matrlx metalloprotelnase lnhlbitor, or a pharmaceutlcally acceptable salt thereof, in admixture with a pharmaceutically acceptable carrler.
Preferably, the matrlx metalloprotelnase lnhlbitor ls a matrlx metalloprotelnase-2 or matrix metalloproteinase-9.
Stlll preferably, the matrlx metalloprotelnase lnhlbltor is a compound selected from the group conslsting of:
3-[[4-(4-fluorophenoxy)benzenesulfonyl]-(1-hydroxycarbamoylcyclopentyl)amlno]proplonlc acld;
4-[4-(4-fluorophenoxy)benzenesulfonylamlno]-CA 022~6303 1998-12-17 tetrahydropyran-4-carboxyllc acld hydroxyamlde;
4-[4-(4-chlorophenoxy)benzenesulfonylmethyl]-tetrahydropyran-4-carboxyllc acid hydroxyamide;
3-[[4-(4-fluorophenoxy)benzenesulfonyl]-(1-hydroxycarbamoylcyclobutyl)amlno]proplonic acld;
4-(4'-chlorobiphenyl-4-yl)-2-[2-(1,3-dioxo-1,3-dihydro-lsolndol-2-yl)ethyl]-4-oxobutyric acid;
{1-[4-(4-fluorobenzyloxy)benzenesulfonyl]-2-hydroxycarbamoylpiperidin-3-yl}carbamic acid isopropyl ester;

2-[4-(4-fluorophenoxy)benzenesulfonylamino]-N-hydroxy-2-methylpropionamide;

3-[4-(4-fluorophenoxy)benzenesulfonyl]-2,N-dlhydroxy-propionamlde;
3-(4-phenoxybenzenesulfonyl)-7-oxa-blcyclo[2.2.1]heptane-2-carboxyllc acld hydroxyamlde;
(4-benzyl-benzyl)-[2-(2,2-dlmethyl-1-methylcarbamoyl-propylcarbamoyl)-4-(4'-fluoroblphenyl-4-yl)butyl]phosphlnic acld;
2-amino-3-[4-(4-fluorophenoxy)benzenesulfonyl]-N-hydroxypropionamide; and N-hydroxy-2-(4-phenylpiperidine-1-sulfonyl)acetamide.
DETAILED DESCRIPTION OF THE INVENTION
Matrix metalloproteinase inhibitors, lncludlng MMP-2 or MMP-9 selectlve lnhibitors, can be prepared according to methods well known to those of ordinary skill ln the art.
Speclfically, matrix metalloproteinase inhibltors with broad CA 022~6303 1998-12-17 - 3a -actlvity and their methods of preparatlon have been described ln PCT Publlcations WO 96/33172, published October 24, 1996, and WO 96/27583, published March 7, 1996; PCT Application No.
PCT/US95/07166, flled June 07, 1995 (which corresponds to United States Patent Application No. 60/017,850, filed May 20, 1996); PCT Application No. PCT/IB95/00427, filed June 6, 1995;
United States Patent Application No. 60/021,652, filed July 12, 1996; United States Patent Application No. 60/024,675, filed August 23, 1996; United States Patent Application No.
60/021,959, filed July 18, 1996; United States Patent Application No. 60/037,600, filed February 11, 1997; Unlted States Patent Appllcation No. 60/037,402, filed February 7, 1997; United States Patent Application No. 60/036,857, filed February 3, 1997; United States Patent Application No.
60/034,535, filed January 6, 1997; European Patent Application 606,046, published July 13, 1994; PCT publlcation WO 90/05719, published May 31, 1990; and European Patent Publication 780,386, published June 25, 1997.

Matrix metallogproteinase -2 or -9 selective int~ ik . ~ refer to compounds thatde",onst~dle a s~le~ity for the MMP-2 or -9 meta ~it~ )~se over MMP-1 meta p.,t ,ase or other MMPs.
The abili~ of matrix meta ~Fruteinase inhibitors or or their pharrnaceut.c~ acceF - !e salts (he.~;nd~r also referred to as the inhibitors of the Fresent invention) to inhibit matrix 10 meta ~Fr~:~,ases or the production of tumor necrosis facbor (TNF) and consequently demonstrate their effectiveness for treating macular degen~dt~n is shown by the following in vitro assay tests.
~ic loqic~l Assav Inhibition of Human Ce'lc ~enase (MMP-1 ) Human ~coi"bi ,ant --'Iz3enase is activated with try~sin using the following ratio: 10 mg trypsin per 100 mg of collagenase. The trypsin and collagenase are incubated at room temperature for 10 minutes then a five fold excess (50 mg/10 mg trypsin) of soybean trypsin inhibitor is added 10 mM stock solutions of inhibitors are made up in dimethyl su*~xide and then diluted usingthef. ~ g Scheme:
10 mM > 120 IlM ~ 12 ~lM--> 1.2 ~,-M--> 0.12 ~lM
Twenty-fi~e microliters of each concer,L,dLion is then added in triplicate to appr~pridte wells of a 96 well mi~rofluor p~ate. The hnal concer,~.Jtion of inhibitor will be a 1:4 dilution after addition of enzyme and substrate. Posit~ve controls (enzyme no inhibitor) are set up in wells D1-D6 and bbnks (no enzyme no inhibitors) are set in wells D7-D12.
Cs'k3enase is diluted to 400 ng/ml and 25 ml is then added to app~upriate wells of the microfluor plate. Final conoerlt~dtion of ~ -3enase in the assay is 100 ng/ml.
Substrate (DNP-Pro-Cha-Gty-Cys(Me) llisA'~ Lys(NMA)-NH2) is made as a 5 mM
stock in dimethyl s~Hoxide and then diluted to 20 mM in assay buffer. The assay is initiated by the addition of 50 ml substrate per well of the microfluor plate to give a hnal concenl,c,tion of 10 mM.
Fluorescen~e readings (360 nM eYciPtion 460 nm efnission) were taken at time 0 and then at 20 minute intervals. The assay is conducted at room ten~pe~ture with a typical assay time of 3 hours.
Fluor~:scenoe vs time is then plotted for both the b~ank and collagenase containing samples (data from triplicate detelll ~dLions is averaged). A bme point that provides a good signal (the blank) and that is on a linear part of the cuNe (usualty around 120 minutes) is chosen to determine ICs" values. The zero time is used as a btank for each compound at each concen~aLon and these values are sul,Oa- lad from the 120 minute data. Data is plotted as inhibitor concentl njon vs % oontrol (inhibitor fluoresc~ce divided by fluorescence of CA 022~6303 1998-12-17 5 C:~c3 nase alone x 100). ICso~s are dete rnined from the concer,t~-tion of inhibitor that gives a signal that is 50% of the control.
If ICso~s are reported to be <0.03 mM then the inhibitors are assayed at conc~nbdtions of 0.3 mM 0.03 mM 0.03 mM and 0.003 mM.
Inhibition of Gelatinase (MMP-2) Inhibition of gel~ti,lase activity is assayed using the Dnp-Pro-Cha41y-Cys(Me) I lis Ala Lys(NMA)-NH2 substrate (10 mM) under the same con 'i;ticns as i~h blticn of human c-'bgenase (MMP-1).
72kD gelatinase is activated with 1 mM APMA (p-aminophenyl mercuric acetate) for 15 hours at 4~C and is diluted to give a final cr~ncen~.,tion in the assay of 100 mg/ml. Inhibitors are diluted as for inh L''icn of human collagenase (MMP-1) to give final conc~r~l-dtions in the assay of 30 mM 3 mM 0.3 mM and 0.03 mM. Each conoen~dlion is done in triplicate.
Fluorescence readings (360 nm p~c~ on 460 emission) are taken at time zero and then at 20 minutes intervals for 4 hours.
ICso's are determined as per inhibition of human ~ g~nase (MMP-1). If ICsols arereported to be less than 0.03 mM then the inhibitors are assayed at final c;oncenl~dtions of 0.3 mM 0.03 mM 0.003 mM and 0.003 mM.
Inhibition of StromelYsin Activity (MMP-3) Inhibition of s~u",elysin activity is based on a modified specbùph~l ."etlic assay descnbed by We;nga, len and Feder (W~ngd~ len H. and Feder J. Spec~ ophoton ,~t, ic Assay for Vertebrate Collagenase Anal. Bioc~em. 147 437 440 (1985)). Hydrolysis of the thio pept~'ide substrate [Ac-Pro-Leu-Gly-SCH[CH2CH(CH3)2~CO-Leu-Gly-OC2Hsl yields a mercaptan fragment that can be i "or.it ~ r~d in the pr~sence of Ellman's reagent.
Human recoi"~ ~ant pruabui,,el~in is a~t,.~l ~ with trypsin using a ratio of 1 ml of a 10 mg/ml trypsin stock per 26 mg of s~ ui "ely~in. The trypsin and stromelysin are ina Ibat~d at 37~C
for 15 minutes followed by 10 ml of 10 mgJml soybean trypsin inhibitor for 10 minutes at 37~C for 10 minutes at 37~C to quench trypsin activity.
Assays are conducted in a total volume of 250 ml of assay buffer (200 mM sodium chloride 50 mM MES and 10 mM cakium chloride pH 6.0) in 96-well microliter plates.
Activated stromelysin is diluted in assay buffer to 25 mg/ml. Ellman's reagent (3-Carboxy4-nitrophenyl disulfide) is made as a 1M stod~ in dimethyl fo~ a~ide and diluted to 5 mM in assay buffer with 50 ml per well yielding at 1 mM hnal conc~nL dtion.
10 mM stock solutions of inhibito~s are made in dimethyl sulfoxide and diluted serially in assay buffer such that addition of 50 mL to the app,uphdte wells yields final conoer~dtions of 3 mM 0.3 mM 0.003 mM and 0.0003 mM. A~l COh 'i~cns are con,plet~ in triplicate.
A 300 mM dimethyl su'~ ide stoclc solution of the peptide substrate is diluted to 15 mM
in assay buffer and the assay is initiated by addition of 50 ml to each well to give a final concer,t,dlion of 3 mM substrate. Blanks c~nsist of the peptide substrate and Ellman's reagent CA 022~6303 1998-12-17 without the enzyme. Product f~""dtion was mont~r~d at 405 nm with a 1~ Devices UVmax plate reader.
IC~ values were det~., n ~ed in the same manner as for c ~ 3 e nase.
Inhibibon of Human 92 kD Celat;.-ase (MMP-9) Inhibition of gelati"ase (MMP-9) activity is assayed using the Mca-Pro-Leu-Gly-Leu-10 Dpa-Ala-Arg-NH2 substrate (10 mM) under similar conditions as des~il,ed above for the inhibition of human e ~'lagenase (MMP-1).
Human recombinant 92 kD gelatinase (MMP-9 gelatinase B) is activated for 2 hourswith 1mM p-aminophenyl-mercuric acetate (from a freshly prepared 100 mM stock in 0.2 N
NaOH) at 37 C.
10 mM dimethyls~ e stock solutions of inhibitors are diluted serially in assay buffer (50 mM TRIS pH 7.5 200 mM NaCI 5 mM CaCI2 20 ~M ZnCI2 0.02% BRIJ-35 (vol./vol.)) using the fc\llowing scheme:
10 mM ) 120 IlM~ 12 ~M ~ 1.2 ~M~ 0.12 IlM
Further dilu~ons are made as necessary following this same scheme. A minimum of four inhibitor concen~dtions for each compound are performed in each assay. 25 ~L of each concenl,dlion is then added to ~i )l cate wells of a black 96 well U-bottomed microfluor plate.
As the final assay volume is 100 llL final conceht,;dlions of inhibitor are the result of a further 14 dilution (i.e. 30 ~M ~ 3 ',lM ~ 0.3 ~M ~ 0.03 ~LM etc.). A blank (no enzyme no inhibitor) and a positive enzyme control (with enzyme no inhibitor) are also prepared in t, iplicdl~.
Activated enzyme is diluted to 100 ng/mL in assay buffer 25 ~L per well is added to appropriate wells of the ",:: oplate. Final enzyme conceh~dtion in the assay is 25 ng/mL
(0.27 nM).
A five mM dimethylsulfoxide stock solution of substrate (Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2) is diluted in assay buffer to 20 ~M. The assay is inrtiated by addition of 50 IlL of diluted substrate yielding a hnal assay concentration of 10 ~lM substrate. A 0 time fluorescenoe reading (320 excitation; 390 emission) is immediately taken and subsequent readings are taken every fifteen minutes at room teinperdt,lre with a PerSeptive Biosystems CytoFluor Multi-Well Plate Reader with the gain at 90 units.
The average value of fluorescence of the enzyme and blank are plotted versus time.
An early bme point on the linear part of this curve is chosen for ICso de~er"~ ,ations. The 0 time point for each compound at each dilution is subl~d- ted from the latter time point and the data then ex~,ressed as percent of enzyme control (inhib~tor fluorescence divided by fluorescenoe of posi~ve enzyme control x 100). Data is plotted as inhibitor concentration versus peroent of enzyme control. ICsos are defined as the concer,~dtion of inhibitor that gives a signal that is 50% of the positive enzyme control.

5Inh.i~''icn of MMP-13 Human r~o",~.,anL MMP-13 is a~vated with 2mM APMA (p-a", ,ophen~l mercuric acetate) for 1.5 hours at 37~C and is diluted to 400 mg/ml in assay buffer (50 mM Tris pH 7.5 200 mM sodium chloride 5mM calcium chbride 20mM zinc chioride 0.02% brij). Twenty-five microliters of diluted enzyme is added per we I of a 96 well microfluor plate. The enzyme is then 10diluted in a 1:4 raUo in the assay by the addition of inhibitor and substrate to givle a final concer,~rdti~n in the assay of 100 mg/ml.
10 mM stock solutions of inhibitors are made up in dimethyl sl ~oYide and then diluted in assay buffer as per the inhibitor dilubon sc~eme for illt,'biti~cn of human c.'lcgenase (MMP-1):
Twenty-five ",:-, :' a of each conce"t,dLon is added in tl r!i ~- to the microfluor pbte. The 15final conoer,t,c Lions in the assay are 30 mM 3mM 0.3 mM and 0.03 mM.
Substrate (Dnp-Pro-Cha~ly-Cys(Me) l lisA'a Lys(NMA)-NH2) is prepared as for inhibition of human l~'k3Pnase (MMP-1) and 50 ml is added to each well to give a final assay concer,t,dtion of 10 mM. Fluor~soence rea.~ings (360 nM Pycit tion; 450 emission) are taken at bme 0 and every 5 minutes for 1 hour.
20Positive controls consist of enzyme and substrate with no inhibitor and blanks consist of substrate only.
ICso's are d~t~.ll. ,ed as per inhit~on of human c~ genase (MMP-1). If ICs0's are reported to be less than 0.03 mM inhibitors are then assayed at final conoenbdtions of 0.3 mM
0.03 mM 0.003 mM and 0.0003 mM.
25All of the compounds of the inventK~n that were tested in the Inhibition of MMP-13 assay had IC50 s of less than 50nm.
Inh blticn of TNF Production The ability of the compounds or ttke pha~ aoeutically accep'-''e salts thereof to inhibit the production of TNF and consequentty d~,nonat dte their effectiveness for treating ~J;~a~s 30involving the producbon of TNF is shown by ffle r,ll~ /~;.,9 in vrtro assay:
Human mononuclear cells were isobted from anti~u'a~~ ~ human blood using a one-step Ficoll-hypaque separation techn ~ue (2) The mononuclear oells were washed three times in Hanks balanced salt solution (HBSS) wffl divalent cations and resuspended to a density of 2 x 106 /ml in HBSS containing 1% BSA Di~r~ntial counts determined using the Abbott Cell Dyn 353500 anatyzer ind'~ t~ that monocytes ranged from 17 to 24% of the total cells in these prepa, dtions.
180m of the oell suspension was ~, t~ into ftat bottom 96 well plates (Costar).
Additions of compounds and LPS (100ng/ml final concent,dLion) gave a final volume of 200ml.
All con~ ns were pe,rl"-"ed in triplicate. After a four hour incubation at 37~C in an humidified 40CO2 incubator plates were removed and oentrifuged (10 minutes at appo,Ai",- 'y 250 x 9) and the supei"atanls removed and assayed for TNFa using the R8D ELISA Kit~.

Inhib;~cn of Anqioqenesis The acbvHy of MMP inhibHors, p.~t~rably MMP-2 or MMP-9 inhibitors, for inhibiting ang egenesis can be determined acco,~ 9 to the t " /,;.~9 in vr~ rat model A 10119 pellet containing an ar,jic3en b material, such as vascular enduU~e'iz' growth factor (VEGF) and a matrix "~ pr~t~ ase inhibitor or pbcebo, is impldl~t ~ intrao~bitally. After one week the eyes are removed and the degree of inhibHion of ans e3enesis is dete~Tnined qualitatively. Inhibitors with IC50's of bss than 10nm reduced angiogenesis. Table 1 de~non~t-.- s that MMP inhibitors are useful in treabng ang :genesis.

Tabk 1 Compound Comeal Neovasculark~,ti3n % Response (#Posi~ve Re~nse~Response) Hydron + PBS 0/3 ~
basic ribrobldal growth factor 3/3 100 3-[14-(4-Fluoro-phenoxy)-benzenesulfonyl]-(1 -hydroxyca~ bai I loyl-cyclopentyl)-amino]-propionic acid 10ng + VEGF 1/4 25 25ng + VEGF 1/6 17 50ng +VEGF 0/5 0 100ng + VEGF 0/5 ~
2 -[4-(4-Fluoro-phenoxy)-benzenesulfonylamino]-N-hydroxy-2-methyl-prip ~na",.de 10ng + VEGF 1/5 20 25ng + VEGF 1/6 17 50ng +VEGF o/5 o 100ng + VEGF 0/5 0 3-(4-Phenoxy-benzenesulfonyl)-7-oxa-bicyclol2.2. 1 Iheptane-2-carboxylic acid hydroxyamide 10ng + VEGF 4/4 25 25ng +VEGF 3/5 60 50ng + VEGF 2/5 40 100ng + VEGF 1/5 20 The c~mposi~ions of the present in~ention may be formulated in a conventional manner using one or more pha""ac~utic~lly ac~eptable carriers. Thus the active compounds of the invenbon may be formulated for oral intraorbital buccal intranasal parenteral ~
10 intravenous intramuscular or subcutaneous) olr rectal administration or in a form suitable for adm ,.~l,dtion by inhalation or insufflation.
The acbve compounds can be a Jnl ,~ ~d in a wide variety of different dosage forms in general the thera~ut,~~~ly effecbve compounds of this invention are present in such dosage fonrns at conc;e"~dLion levels ranging from about 5.0% to about 70% by weight.

For oral aJ~" ,i~t,d~n bblets containing various a.- i~ -nt~ such as microcrystalline cellulose sodium citrate cakium cdlt~ondle dicalcium phosph3t~ and glyane may be employed along with vanous disintegrants such as starch (and pr~:ferdb~J com potato or tapioca starch)~
alginic acid and oertain complex silicates together with granulation binders like polyvinylpyrrolidone sucrose gebtion and acacia. Ad~'i'icna'ly lubricat,ng agents such as r"agnesium stearate sodium buryl sulfate and talc are often very useful for IdbleUing purposes.
Solid compositions of a simibr type may also be employed as fillers in gelatin ~psu~es; p,ef~ d ",ateria.~ in this conne- lion also include lactose or milk sugar as well as high ",~'ecul-~ weight polyethylene glycols. When aqueous suspensions and/or elixirs are desired for oral ad", ~i~t,ation the ac~ve i"y~ -nt may be combined with vanous s.~eetel, -g or flavoring agents colonng matter or dyes and if so desired emulsifying andJor suspending agents as well together wtth such diluents as water ethanol propylene glycol glycerin and various like coi"~:nalions thereof. In the case of animals they are advantageously contained in an animal feed or drinking water in a concentldlion of 5-5000 ppm preferably 25 to 500 ppm.
For intraorbital ad" ;ni~.tiation a sterile injectable solution of the active ingredient is usually prepared. Solutions of a therapeutic compound of the present invention in an aqueous solution or suspension (particle size less than 10 micron) may be employed. The aqueous solutions should be suitably adjusted and buffered pre:f~rably at a pH between 5 and 8 if necessary and the liquid diluent first r~nder~d isotonic. Small amounts of polymers can be added to increase viscosity or for sustained release (such as cellulosic polymers Dextran polyethylene glycol or alginic ~id). These solutions are suitable for i,l~,aoibital injection purposes. The prepald~on of all these solutions under stenle cor,dilions is readily accomplished by sldnda,d pha""aoeutic~l techn.~ues well known to those skilled in the art. In the case of animals compounds can be a-ll", tered intraorbitally at dosage levels of about 0.1 to 50 mgJkg/day advdr'~geously 0.2 to 10 mglkg/day given in a single dose or up to 3 divided doses.
For pa(t:"t dl adrr, ,;~tidtion (intramuscular i"~dpe"toneal subcutaneous and intravenous use) a sterile i"je t- solution of the active ingr~ 'ienl is usually prepared.
Solutions of a therapeu~c compound of the present invention in either sesame or peanut oil or in aqueous propylene gly~ol may be employed. The aqueous solutions should be suitably adjusted and buffered prefeldbly at a pH of greater than 8 if necessary and the liquid diluent first rendered isotonic. These aqueous solutions are suitable for intravenous injection purposes. The oily solutions are suitabb for i"t,da,Licular intramuscular and subcutaneous injection purposes.
The p,~:par.~t~n of all these solutions under sterile con iticns is readily ac~",p'i~,hed by s~dnda~d pha",.aceut~l te~l,r. !es well known to those skilled in the art. In the case of animals compounds c~n be admi., : ed intramuscularly or subcutaneously at dosage levels of about 0.1 to 50 mg/kg/day advar~geously 0.2 to 10 mg/kg/day given in a single dose or up to 3 divided doses.

CA 022~6303 1998-12-17 The actlve compounds may also be formulated in rectal compositions such as suppositories or retention enemas, e.q., contalnlng conventlonal suppository bases such as cocoa butter or other glycerides.
For intranasal administration or adminlstration by inhalatlon, the active compounds are conveniently delivered in the form of a solutlon or suspension from a pump spray container that is squeezed or pumped by the patient or as an aerosol spray presentation from a pressurized container or a nebulizer, with the use of a suitable propellant, e.q., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol, the dosage unit may be determined by providlng a valve to deliver a metered amount. The pressurized contalner or nebulizer may contain a solution or suspension of the active compound. Capsules and cartridges (made, for example, from gelatln) for use ln an inhaler or insufflator may be formulated containing a powder mix of a compound of the invention and a suitable powder base such as lactose or starch.
The pharmaceutical composition of this invention may be put in a commercial package for practical use, storage and transportation. Such a commercial package usually carries a written matter which describes that the pharmaceutical composition can or should be used for the purpose described in this specification.

Claims (13)

1. A pharmaceutical composition for treating or preventing ocular angiogenesis by inhibiting a matrix metalloproteinase in a mammal, which comprises (a) a matrix metalloproteinase inhibiting effective amount of a matrix metalloproteinase inhibitor, or a pharmaceutically acceptable salt thereof and (b) a pharmaceutically acceptable carrier.

2. A pharmaceutical composition according to claim 1, wherein the matrix metalloproteinase inhibitor is a matrix metalloproteinase-2 or matrix metalloproteinase-9 inhibitor.

3. A pharmaceutical composition according to claim 1, wherein the matrix metalloproteinase inhibitor is a matrix metalloproteinase-2 selective or matrix metalloproteinase-9 selective inhibitor.

4. A pharmaceutical composition according to claim 1, wherein the matrix metalloproteinase inhibitor is a compound selected from the group consisting of:
3-[[4-(4-fluorophenoxy)benzenesulfonyl]-(1-hydroxycarbamoylcyclopentyl)amino]propionic acid;
4-[4-(4-fluorophenoxy)benzenesulfonylamino]tetrahydro-pyran-4-carboxylic acid hydroxyamide;
4-[4-(4-chlorophenoxy)benzenesulfonylmethyl]tetrahydro-pyran-4-carboxylic acid hydroxyamide;
3-[[4-(4-fluorophenoxy)benzenesulfonyl]-(1-hydroxy-carbamoylcyclobutyl)amino]propionic acid;
4-(4'-chlorobiphenyl-4-yl)-2-[2-(1,3-dioxo-1,3-dihydroisoindol-2-yl)ethyl]-4-oxobutyric acid;
{1-[4-(4-fluorobenzyloxy)benzenesulfonyl]-2-hydroxy-carbamoylpiperidin-3-yl}-carbamic acid isopropyl ester;
2-[4-(4-fluorophenoxy)benzenesulfonylamino]-N-hydroxy-2-methylpropionamide;
3-[4-(4-fluorophenoxy)benzenesulfonyl]-2,N-dihydroxy-propionamide;
3-(4-phenoxybenzenesulfonyl)-7-oxabicyclo[2.2.1]heptane-2-carboxylic acid hydroxyamide;
(4-benzylbenzyl)-[2-(2,2-dimethyl-1-methylcarbamoylpropyl-carbamoyl)-4-(4'-fluoro-biphenyl-4-yl)-butyl]phosphinic acid;
2-amino-3-[4-(4-fluorophenoxy)benzenesulfonyl]-N-hydroxypropionamide; and N-hydroxy-2-(4-phenylpiperidine-1-sulfonyl)acetamide.

5. A pharmaceutical composition according to claim 1, wherein the matrix metalloproteinase inhibitor is 3-[[4-(4-fluorophenoxy)benzenesulfonyl]-(1-hydroxycarbamoyl-cyclopentyl)amino]propionic acid.

6. A pharmaceutical composition according to claim 1, wherein the matrix metalloproteinase inhibitor is 2-[4-(4-fluorophenoxy)benzenesulfonylamino]-N-hydroxy-2-methyl-propionamide.

7. A pharmaceutical composition according to claim 1, wherein the matrix metalloproteinase inhibitor is 3-(4-phenoxybenzenesulfonyl)-7-oxabicyclo[2.2.1]heptane-2-carboxylic acid hydroxyamide.

8. A pharmaceutical composition according to any one of claims 1 to 7, adapted for use in a human.

9. A pharmaceutical composition according to any one of claims 1 to 8, which is an intraorbital composition.

10. A pharmaceutical composition according to any one of claims 1 to 9, wherein the matrix metalloproteinase inhibitor is contained at a dosage of 0.2 to 10 mg/kg/day.

11. A commercial package comprising the pharmaceutical composition of any one of claims 1 to 10 and a written matter describing that the pharmaceutical composition can or should be used for treating or preventing ocular angiogenesis.

12. A commercial package according to claim 11, wherein the written matter describes that the pharmaceutical composition can or should be used for treating diabetic retinopathy.

13. A commercial package according to claim 11, wherein the written matter describes that the pharmaceutical composition can or should be used for treating age-related macular degeneration (ARMD).